Essential oil composition of three accessions of Dracocephalum heterophyllum Benth. cultivated at Palampur, India

Seeds of three accessions of Dracocephalum heterophyllum growing wild in high altitude locations in the cold desert regions of trans-Himalaya were collected and propagated under polyhouse conditions at IHBT Palampur to evaluate, compare their growth performance and essential oil composition. Three different chemotypes viz., citronellol-rose oxides type, citronellol type and citronellal-trans rose oxide type were identified. Samples of essential oils were distilled separately from leaves and inflorescence of all the three cultivated accessions and characterised by capillary GC and GC-MS techniques, which showed variation in the volatile constituents. In all the three accessions, citronellol was higher in the inflorescence oils as compared to leaf oils. Rose oxides, which are important high odour value cyclic monoterpene ethers were found in the essential oil samples of leaves as well as in the inflorescence of accession II, whereas these constituents were absent in the leaves of other two accessions. The essential oil from the inflorescence part of accession I was devoid of both cis and trans isomers of rose oxide, while accession III contained only one isomer of trans rose oxide. Under polyhouse conditions, seed germination was higher in accession III, whereas aerial biomass was higher in accession I. In comparison to other accessions, yield of volatile oil was higher in both leaf and inflorescence of accessions I, whereas accession II had higher oil yield from inflorescence. This is the first report of volatile oil composition of three accessions of D. heterophyllum cultivated at Palampur having potential as a new source of high valued essential oil for introduction in the perfumery industry.     

Authentication and quantitative analysis on the chemical profile of Xanthium fruit (Cang-Er-Zi) by high-performance liquid chromatography-diode-array detection tandem mass spectrometry method

A high-performance liquid chromatographic method using diode-array detection and electrospray ionization tandem mass spectrometry (HPLC-DAD-ESI-MS/MS) was developed for the qualitative and quantitative analysis on Xanthium fruit, a commonly used traditional Chinese medicine. In this study, 7 characteristic components, 1-O-caffeoylquinic acid, 4-O-caffeoylquinic acid, chlorogenic acid, 1,5-O-di-caffeoylquinic acid, 1,3-O-di-caffeoylquinic acid, 4,5-O-di-caffeoylquinic acid and 1,3,5-O-tri-caffeoylquinic acid were identified and quantified by a validated HPLC-DAD method, and a fingerprint comprised of 12 markers was established under the same operating conditions. Furthermore, HPLC-ESI-MS/MS method was successfully used to deduce the structure of three main constituents. On the basis of the established chromatographic profiles, 30 populations of cocklebur samples including 3 related species and 1 unknown species were divided into 3 chemotypes, indicated that place of origin significantly influences the kinds and content of components in cocklebur, and hence affects their quality. The simultaneous determination of 7 caffeoylquinic acids in the 30 samples showed a great variety in the amounts of caffeoylquinic acids present. The study indicated that some species such as Xanthium mongolicum of the genus Xanthium might be suitable for development as new alternative sources of caffeoylquinic acids to supplement the officially listed Xanthium species, and the abundant constituents such as chlorogenic acid perhaps should be recorded in some authorized publications and applied to the quality control or quality evaluation for Xanthium in China. The entire analytical procedure is reproducible and suitable for the authentication and quantification of Xanthium fruits.     

Lipids of elaeagnus fruit

The composition of the classes of lipids of the fruit of three morphological forms of the elaeagnusElaeagnus angustifolia L. have been studied. Their compositions were identical. The amounts of the main lipid classes of extracts of the seeds and pericarps, the fatty acid compositions of the acyl-containing classes of lipids, and the compositions of the carbohydrates and sterols have been determined. The fatty oil of the seeds contained linoleic acid, while the main fatty acids of the pericarp extracts were the 16:0, 18:1, and 18:2 acids. The 16:1 acid that is characteristic for sea buckthorn oil was detected in the elaegnus fruit in insignificant amounts. The class of sterols, both in the free and in the esterified states, was represented by -sitosterol. The main hydrocarbon of the pericarps and seeds was nonacosane.Institute of the Chemistry of Plant Substances, Academy of Sciences of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 17–21, January–February, 1990.     

The Intehaction of Aqueous Solutions of Chlorine with Malic Acid,Tartaric Acid,and Various Fruit Juices: A Source of Mutagens

Abstract

The interactions of aqueous solutions of chlorine with some fruit acids (citric acid, DL-malic acid, arid L-tartaric acid) at different pH values were studied. Diethyl ether extraction followed by GC/MS analysis indicated that a number of mutagens (certain chlorinated propanones and chloral hydrate) are present as major products in some of these samples. A number of fruit juices (orange, grape, apple, pineapple, and grapefruit) were also treated with aqueous solutions of chlorine at their pH values. The products were analyzed by GC/MS. The same mutagens that were formed by the pure acids (citric acid and DL-malic acid) were identified as major products in ether extracts of these samples. Another mutagen, dichloroacetonitrile, was also identified as a minor product in some of these juice samples. All of the major products observed in the chlorination of all five fruit juices are potentially derived from reactions of aqueous solutions of chlorine with citric or malic acid and with trace amounts of acetaldehyde and acetone in the juices. The minor product, dichloroacetonitrile, is likely derived from the chlorination of certain amino acids in the fruit juices.     

Rapid automated high performance liquid chromatography method for simultaneous determination of amino acids and biogenic amines in wine, fruit and honey

This paper reports a new, simple, rapid and economical method for routine determination of 24 amino acids and biogenic amines in grapes and wine. No sample clean-up is required and total run time including column re-equilibration is less than 40min. Following automated in-loop automated pre-column derivatisation with an o-phthaldialdehyde, N-acetyl-l-cysteine reagent, compounds were separated on a 3mm×25cm C(18) column using a binary mobile phase. The method was validated in the range 0.25-10mg/l; repeatability was less than 3% RSD and the intermediate precision ranged from 2 to 7% RSD. The method was shown to be linear by the 'lack of fit' test and the accuracy was between 97 and 101%. The LLOQ varied between 10μg/l for aspartic and glutamic acids, ethanolamine and GABA, and 100μg/l for tyrosine, phenylalanine, putrescine and cadaverine. The method was applied to grapes, white wine, red wine, honey and three species of physalis fruit. Grapes and physalis fruit were crushed, sieved, centrifuged and diluted 1/20 and 1/100, respectively, for analysis; wines and honeys were simply diluted 10-fold. It was shown using this method that the amino acid content of grapes was strongly correlated with berry volume, moderately correlated with sugar concentration and inversely correlated with total acidity.     

Use of an Optimum Wavelength to Detect Glutamic Acid and its Metabolites in Human Serum by Reversed-Phase HPLC

Abstract

Dansylated glutamic acid, glutamine and γ-amino butyric acid (GABA) show maximum absorption at 221 nm. Using this wavelength, the detection limits for dansylated amino acids studied by reversed-phase HPLC are similar to those reported by fluorescence. This technique was used to look foe the presence of glutamic acid and its metabolites in human serum. Glutamic acid and glutamine were present in significant amounts and their levels were 2.5 and 6.1 nmoles/ml respetively, while GABA was present in trace amounts, less than 0.3 nmoles/ml.     

气相色谱-质谱测定樟林番荔枝种子挥发油的脂肪酸组成

用溶剂萃取法提取樟林番荔枝果实种子中的挥发性物质,测定出其挥发油质量分数为13.3%;利用GC-MS方法分离确认出其中的9种化学成分;用面积归一化法得出了9种脂肪酸在挥发油中的质量分数;其中9-十八烯酸占49.42%,十六酸占20.37%,十八酸占14.16%,9,2-十八二烯酸占13.59%;不饱和脂肪酸,占63.01%.该项研究给番荔枝果实的深入开发利用及种质资源的有效保护提供了科学依据.     

柽柳实中挥发油和脂肪酸分析

首次研究了维药细穗柽柳(Tamarix leptostachys Bunge)实中挥发油和脂肪酸的化学成分。 分别采用药典中的挥发油提取法和索式取提法提取柽柳实中的挥发油和脂肪酸,使用气质联用技术获取总离子流图,各色谱峰相应的质谱图经过NIST2011标准谱库检索定性,并采用峰面积归一化法进行定量分析,计算各成分的相对百分含量。 两种方法分别鉴定出48种挥发油和19种脂肪酸。 挥发油主要成分为芳香类化合物(43.71%)、芳香性醛酮类(20.58%)、脂肪酸类(13.03%)、酯类(17.36%)和醇类(4.19%)等。 脂肪酸主要成分为棕榈酸(35.61%)、亚油酸(27.26%)和油酸(11.33%)等,其中不饱和脂肪酸含量占总脂肪酸含量的38.65%。 维药细穗柽柳实中富含丰富的挥发油和不饱和脂肪酸,具有很好的开发利用价值。     

A first glance into the nutritional properties of the sea cucumber Parastichopus regalis from the Mediterranean Sea (SE Spain)

This work reports for the first time the nutritional profile, including proximate chemical composition, amino acids, fatty acids and minerals of Parastichopus regalis from the Mediterranean Sea (SE Spain). The studied species had a high moisture content, moderate protein and low lipid levels. The most abundant amino acids were glutamic acid, arginine and tyrosine. Polyunsaturated fatty acids, especially arachidonic acid, dominated the fatty acid profile. Iron, sodium, calcium and zinc were the most abundant mine rals. In general, P. regalis has a balanced nutritional quality suitable for human consumption.     

Robust and straightforward chemo-enzymatic enantiopure dipeptide syntheses and diketopiperazines thereof

We have explored the scope of the synthetic route towards d-phenylglycyl diketopiperazines, involving a penicillin acylase catalysed formation of d-phenylglycyl dipeptides of l-amino acids with functional groups in the side chain. The synthesis of dipeptides from serine, threonine, glutamic acid, glutamine and methionine was successful. In contrast, aspartic acid, asparagine and cysteine only afforded trace amounts of dipeptides while no dipeptide was detected with arginine, lysine and tyrosine. Isolated dipeptide yields varied from 10% to 76%. The dipeptides were successfully converted into their corresponding enantiopure diketopiperazines by chemical esterification and cyclization under alkaline conditions, in 35–43% yield. In the case of glutamic acid, the procedure yielded the diketopiperazine with an esterified side chain. Remarkably with glutamine, the amide function in the side chain was transformed into an ester moiety, resulting in the same diketopiperazine as with glutamic acid.     

Use of reversed-phase high-performance liquid chromatography for simultaneous determination of glutamine synthetase and glutamic acid decarboxylase in crude extracts

Glutamine and gamma-aminobutyric acid (GABA), formed from glutamic acid in crude tissue extracts by glutamine synthetase and glutamic acid decarboxylase respectively, were separated by derivatization with dansyl chloride followed by reversed-phase high-performance liquid chromatography on the Altex Ultrasphere ODS-5 column. The mobile phase was a gradient of 100 mM potassium dihydrogen phosphate (pH 2.1) with 0-40% acetonitrile. The amounts of glutamine and GABA formed from glutamic acid were determined under different reaction conditions.     

Application of stepwise discriminant analysis to classify commercial orange juices using chiral micellar electrokinetic chromatography-laser induced fluorescence data of amino acids

The use of chiral amino acids content and stepwise discriminant analysis to classify three types of commercial orange juices (i.e., nectars, orange juices reconstituted from concentrates, and pasteurized orange juices not from concentrates) is presented. Micellar electrokinetic chromatography with laser-induced fluorescence (MEKC-LIF) and beta-cyclodextrins are used to determine L- and D-amino acids previously derivatized with fluorescein isothiocyanate (FITC). This chiral MEKC-LIF procedure is easy to implement and provides information about the main amino acids content in orange juices (i.e., L-proline; L-aspartic acid, D-Asp, L-serine, L-asparagine, L-glutamic acid, D-Glu, L-alanine, L-.arginine, D-Arg, and the non-chiral gamma-amino-n-butyric acid (GABA), i.e., gamma-aminobutyric acid). From these results, it is clearly demonstrated that some D-amino acids occur naturally in orange juices. Application of stepwise discriminant analysis to 26 standard samples showed that the amino acids L-Arg, L-Asp and GABA were the most important variables to differentiate the three groups of samples. With these three selected amino acids a 100% correct classification of the samples was obtained either by standard or by leave-one-out cross-validation procedures. These classification functions based on the content in L-Arg, L-Asp and GABA were also applied to nine test samples and provided an adequate classification and/or interesting information on these samples. It is concluded that chiral MEKC-LIF analysis of amino acids and stepwise discriminant analysis can be used as a consistent procedure to classify commercial orange juices providing useful information about their quality and processing. To our knowledge, this is the first report about the combined use of chiral capillary electrophoresis and discriminant techniques to classify foods.     

Husk Cherry: Nutritional attributes,bioactive compounds and technological applications

Husk cherry (Physalis pruinosa; L) is a tropical fruit commonly grown in the Mediterranean region, and widely consumed fresh in most countries of the world. To enhance our knowledge about this promising fruit, the technological, therapeutic, and nutritional quality attributes of the raw fruits were investigated. The fruit had an orange yellowish color and a mild sweetness, with a pulp yield of 93.40 % (w/w) and a juice yield of 64.45 % (v/w). It contains high ratios of total sugar (41.65%), protein (14.56%), crude fibers (11.59 %), ascorbic acid (40.17 mg/100 g), ß-carotene (64.78 μg/g), total phenols (178.53 mg gallic acid 100 g⁻¹), anthocyanin (7.01 μg cyaniding-3-glucoside equivalents 100 g⁻¹), flavonoids (6.69 mg quercetin g⁻¹), and chlorophylls (19.03 μg g⁻¹). These fruit showed high antioxidant activities, which were found using DPPH and ABTS assays, and could be considered as a good source of antioxidants. The fruit contained several minerals, including potassium, phosphorus, sodium, and magnesium. The major amino acids in the fruit pulp were glutamic acid, aspartic acid, arginine, phenylalanine, proline, valine and alanine. The main polyphenols detected in the fruits were gallic acid, 4-hydroxybenzoic acid, benzoic acid, catechol, ellagic acid, chlorogenic acid, and epicatechin. Several flavonoids can be obtained in high concentrations, such as naringenin, luteolin, and rutin. Six functional food products (juice, jelly, jam, candied fruits, chocolate--coated dried fruits, and fruit leather) were manufactured using husk cherry fruits. The quality attributes of prepared products were determined. The highest crude fiber, ascorbic acid, β carotene, and total sugar compositions were found in chocolate-coated dry fruit products. All the manufactured products were well palatable amongst 25 trained panelists. The findings of this study confirmed the nutritional and technological potential of husk cherry fruits, which might be used effectively within the development of some functional products for food industries.     

Discrimination of Genetically Very Close Accessions of Sweet Orange (Citrus sinensis L. Osbeck) by Laser-Induced Breakdown Spectroscopy (LIBS)

The correct recognition of sweet orange (Citrus sinensis L. Osbeck) variety accessions at the nursery stage of growth is a challenge for the productive sector as they do not show any difference in phenotype traits. Furthermore, there is no DNA marker able to distinguish orange accessions within a variety due to their narrow genetic trace. As different combinations of canopy and rootstock affect the uptake of elements from soil, each accession features a typical elemental concentration in the leaves. Thus, the main aim of this work was to analyze two sets of ten different accessions of very close genetic characters of three varieties of fresh citrus leaves at the nursery stage of growth by measuring the differences in elemental concentration by laser-induced breakdown spectroscopy (LIBS). The accessions were discriminated by both principal component analysis (PCA) and a classifier based on the combination of classification via regression (CVR) and partial least square regression (PLSR) models, which used the elemental concentrations measured by LIBS as input data. A correct classification of 95.1% and 80.96% was achieved, respectively, for set 1 and set 2. These results showed that LIBS is a valuable technique to discriminate among citrus accessions, which can be applied in the productive sector as an excellent cost–benefit tool in citrus breeding programs.     

Gas chromatographic-mass spectrometric fragmentation study of flavonoids as their trimethylsilyl derivatives: analysis of flavonoids, sugars, carboxylic and amino acids in model systems and in citrus fruits

The fragmentation patterns and quantitation possibilities of three anthocyanidins (pelargonidin, cyanidin, malvidin), one flavonol (quercetin), two flavones (apigenin, luteolin) and two flavanones (naringenin, hesperetin) have been investigated as trimethylsilyl and as trimethylsilyl (oxime) derivatives by gas chromatography-mass spectrometry. Results proved that anthocyanidins and flavanones form trimethylsilyl (oximes), while flavonol and flavones provide simple trimethylsilyl derivatives. In all cases, characteristic fragments of high masses are formed proper for quantitation purposes. Hydrolysis conditions for naringin, hesperidin and rutin have been optimized, resulting in the quantitative release of naringenin, hesperetin and quercetin together with their corresponding saccharides. These basic studies made possible the identification and quantification of the flavonoid, carboxylic-/amino acid and sugar constituents of citrus fruit juices and albedos, without any extraction/enrichment procedure. In total 33 compounds have been determined in hydrolyzed samples, such as 2 flavonoids (naringenin and hesperetin), 6 phenolic acids (trimethoxybenzoic, 4-hydroxybenzoic, vanillic, quinic, chlorogenic and rosmarinic acids), 3 aliphatic carboxylic acids (levulinic, malic, citric acids), phosphoric acid, 4 amino acids (aspartic, glutamic acids, alanine, proline), 9 monosaccharides (xylose, arabinose, rhamnose, fucose, fructose, galactose, glucose, galacturonic acid, sedoheptulose), inositol, sugarphosphate, 5 disaccharides and tocopherol. Measurements were carried out as the trimethylsilyl (oxime) ether/ester derivatives of constituents, in the concentration range of 2 x 10(-3) to 49.9%. Identification level of samples varied between 26.4 and 77.5%, expressed in dry matter content of juices and albedos.     

Lipids of the fruit of Diospyros kaki

The composition and amounts of the various groups of liposoluble compounds in the rind and flesh of persimmon fruit of the varieties Hachiya and Hyakume have been established by a combination of chromatographic and chemical methods. The identity of the qualitative composition of the lipids of the flesh and rind of the fruit and quantitative differences in the amounts of individual groups have been found. A total of 24 groups of compounds were identified, the main ones of which were monogalactosyldiglycerides, carotenoids, diacylglycerols, ceramide oligosides, digalactosyldiglycerides, phosphatidylglycerols; phosphatidylcholines, and free and glycosylated sterols. In the fatty acid composition of the lipids unsaturated fatty acids — linolenic, oleic, linoleic, and palmitoleic — predominated (>70%).     

Variations in chemical fingerprints and major flavonoid contents from the leaves of thirty‐one accessions of Hibiscus sabdariffa L.

The leaves of Hibiscus sabdariffa L. have been used as traditional folk medicines for treating high blood pressure and fever. There are many accessions of H. sabdariffa L. throughout the world. To assess the chemical variations of 31 different accessions of H. sabdariffa L., fingerprinting analysis and quantitation of major flavonoids were performed by high‐performance liquid chromatography (HPLC). The HPLC method was validated for linearity, sensitivity, precision, repeatability and accuracy. A quadrupole‐time‐of‐flight mass spectrometry (Q‐TOF‐MS) was applied for the characterization of major compounds. A total of 9 compounds were identified, including 6 flavonoids and 3 phenolic acids. In the fingerprint analysis, similarity analysis (SA) and principal component analysis (PCA) were used to differentiate the 31 accessions of H. sabdariffa L. Based on the results of PCA and SA, the samples No. 15 and 19 appeared much different from the main group. The total content of five flavonoids varied greatly among different accessions, ranging from 3.35 to 23.30 mg/g. Rutin was found to be the dominant compound and the content of rutin could contribute to chemical variations among different accessions. This study was helpful to understand the chemical variations between different accessions of H. sabdariffa L., which could be used for quality control. © 2015 The Authors Biomedical Chromatography Published by John Wiley & Sons Ltd.     

Determination of Pentacyclic Triterpenes from Betula utilis by High-Performance Liquid Chromatography and High-Resolution Magic Angle Spinning Nuclear Magnetic Resonance Spectroscopy

A rapid and reliable method was developed and validated for determining betulin and betulinic acid in bark in Betula utilis by high-resolution magic angle spinning ¹H nuclear magnetic resonance (HR-MAS NMR) spectroscopy. HR-MAS NMR spectroscopy clearly distinguished the resonances of betulin and betulinic acid in the bark of all accessions of B. utilis. The concentrations of betulin and betulinic acid were calculated and added to the spectra. The determination of the targeted metabolites in chloroform extract of bark of each accession of B. utilis was performed by high-performance liquid chromatography (HPLC). Quantitatively, betulin was present at higher concentrations than betulinic acid in all accessions. The HR-MAS NMR and HPLC results showed that betulin and betulinic acid varied significantly among accessions of B. utilis. Principal component analysis of the NMR and HPLC results provided classification into three metabolic groups in which the betulin concentration was high, moderate, or low. The results show that HR-MAS NMR is rapid for fingerprinting of betulin and betulinic acid in the bark of B. utilis, while minimizing the drawbacks associated with solvent extraction.     

Effect of additives on the amination of 2-oxoglutaric acid by gamma-radiation

The effect of added substances was studied on the yield of glutamic acid produced by gamma-ray irradiation of 2-oxoglutaric acid and ammonia in aqueous solution. The contents of amino acids in the irradiated solutions were determined with amino acids analyzer. Sodium nitrate, allyl alcohol or sodium formate was used as an added substance. The yield of glutamic acid significantly decreased by the addition of nitrate, and it was little affected by the addition of allyl alcohol. In the presence of formate the yield increased from G = 0.4 (2-oxoglutaric acid 0.05M and ammonium hydroxide 2M) to G = 1.1. As a result, it was found that hydrated electron contributes on the formation of glutamic acid, but hydroxyl radical does not. The yield showed a maximum at ca. 0.1 M ammonium hydroxide concentration. These facts indicate that NH2 radical does not contribute to the formation of glutamic acid. As a reaction mechanism, it can be explained that 2-oxoglutaric acid which had been reduced by hydrated electron reacts with ammonia.     

Synthesis of novel fullerene α-amino acid conjugates

Aspartic acid and glutamic acid with protected α-amino and α-carboxyl groups had been used to react with the activated hydroxyl group of N-substituted 3,4-fullero pyrrolidine. The products were deprotected, affording two monofuUerene α-amino acids, monofullerene aspartic acid （mFas） and monofullerene glutamic acid （mFgu）. Then a bifullerene glutamic acid conjugate （bFguC） was synthesized by reaction of mFgu containing protected amino group with N-substituted 3,4-fullero pyrrolidine.